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Wenbo Zhang Xuemei Xu Raymond Kao Tina Mele Peter Kvietys Claudio M. Martin Tao Rui 《PloS one》2014,9(9)
Myocardial contractile dysfunction in sepsis is associated with the increased morbidity and mortality. Although the underlying mechanisms of the cardiac depression have not been fully elucidated, an exaggerated inflammatory response is believed to be responsible. Nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) inflammasome is an intracellular platform that is involved in the maturation and release of interleukin (IL)-1β. The aim of the present study is to evaluate whether sepsis activates NLRP3 inflammasome/caspase-1/IL-1β pathway in cardiac fibroblasts (CFs) and whether this cytokine can subsequently impact the function of cardiomyocytes (cardiac fibroblast-myocyte cross-talk). We show that treatment of CFs with lipopolysaccharide (LPS) induces upregulation of NLRP3, activation of caspase-1, as well as the maturation (activation) and release of IL-1β. In addition, the genetic (small interfering ribonucleic acid [siRNA]) and pharmacological (glyburide) inhibition of the NLRP3 inflammasome in CFs can block this signaling pathway. Furthermore, the inhibition of the NLRP3 inflammasome in cardiac fibroblasts ameliorated the ability of LPS-chalenged CFs to impact cardiomyocyte function as assessed by intracellular cyclic adenosine monophosphate (cAMP) responses in cardiomyocytes. Salient features of this the NLP3 inflammasome/ caspase-1 pathway were confirmed in in vivo models of endotoxemia/sepsis. We found that inhibition of the NLRP3 inflammasome attenuated myocardial dysfunction in mice with LPS and increased the survival rate in mice with feces-induced peritonitis. Our results indicate that the activation of the NLRP3 inflammasome in cardiac fibroblasts is pivotal in the induction of myocardial dysfunction in sepsis. 相似文献
33.
Relationships between induced high leaf intercellular CO2 concentrations, leaf K+ and NO3 ? ion movement and early fruit formation under macronutrient limitation are not well understood. We examined the effects and interactions of reduced K/N input treatments on leaf intercellular CO2, photosynthesis rate, carboxylation and water use efficiency, berry formation as well as leaf/fruit K+, NO3 ? and photosynthate retention of strawberry (Fragaria × ananassa Duch.) to enhance low-input agriculture. The field study was conducted in Nova Scotia, eastern Canada during 2009–2010. The experimental treatments consisted of five K2O rates (0, 6, 12, 18, and 24 kg ha?1) and five N rates (0, 5, 10, 15, and 20 kg ha?1), representing respectively, 0, 25, 50, 75, and 100 % of regular macronutrient recommendations based on the soil testing. The treatments were arranged in a split-plot design with three blocks in the field. The cultivar was ‘Mira’, a June-bearing crop. The results showed that strawberry plants treated with 25 %-reduced inputs could induce significantly higher leaf intercellular CO2 concentrations to improve plant photosynthesis, carboxylation and water use efficiency and translocation of leaf/fruit K+ and dissolved solids, which could advance berry formation by 6 days and produce significantly higher marketable yields (P < 0.05). Higher leaf intercellular CO2 inhibited leaf/fruit NO3 ? ion retention, but this inhibition did not occur in leaf/fruit K+ retention. Linear interactions of the K/N treatments were significant on fruit marketable yields, intercellular CO2, net photosynthesis, leaf transpiration rates, and leaf temperatures (P < 0.05). It was concluded that higher leaf CO2 could enhance plant photosynthesis, promote plant carboxylation and water use efficiency, and advance berry formation, but it could inhibit leaf NO3 ? retention. This inhibition did not find in leaf K+ ion and dissolved solid retention. Overlay co-limitation of leaf intercellular CO2 and translocation of leaf/fruit K+/NO3 ? and total dissolved solids could constrain more fruit formation attributes under full macronutrient supply than reduced inputs. It was suggested that low input would be an optimal and sustainable option for improving small fruit crop physiological development and dealing with macronutrient deficiency challenge. 相似文献
34.
We used a N-biotinylated peptide analog of the C-terminal domain of the tumor suppressor protein, p21cip1/waf1 to elucidate peptide/protein interacting partners. The C-terminal domain of p21cip1/waf1 protein spanning 141-160 amino acid residues is known to bind PCNA and this interaction is important in many biological processes including cell-cycle control. This C-terminal 20-mer efficiently extracts PCNA in the presence of a variety of N- or C-terminally attached affinity tags. Using difference silver stained 2D gels combined with in-gel tryptic digests, we identified the difference spots using MALDI-TOF mass spectrometry-based peptide mass fingerprinting followed by a database search using PROFOUND against NCBIs human nonredundant protein sequence data bank. Identified spots include the p48 subunit of chromatin assembly factor-1, the heat shock 70 protein analog BiP, calmodulin, nucleolin and a spot similar in size to dimeric PCNA. In contrast, microcapillary ion-trap LC-MS/MS analysis of a tryptic digest of entire affinity extracts derived from both control and experimental runs followed by database searches using SEQUEST confirmed the presence of most of the above proteins. This strategy also identified hnRNPA1, HPSP90alpha, HSP40 and T-complex protein 1, a protein similar to prothymosin, and a possible allelic variant of the p21cip1/waf1 protein. The use of N-biotinylated peptide derived from the C-terminal domain of p21cip1/waf1 protein in proteomic analysis exemplified here suggests that peptides obtained from intracellular functional screens could also potentially serve as efficient baits to discover new drug targets. 相似文献
35.
A cDNA library was prepared from poly(A) mRNA extracted from 9-day hamster-yolk-sac erythroid cells. Two clones containing inserts coding for embryonic beta-like z or y globin-chains were isolated. Their identity was confirmed by (a) translation of hybrid selected mRNAs and (b) nucleotide sequence analysis of the inserts and comparison to the embryonic beta-like globin genes of Balb/c mouse. Availability of sequences for embryonic and adult globin cDNAs will aid in investigations of the molecular mechanisms of the globin switch in hamster YSEC. 相似文献
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Hailong Jin Congran Li Ding Li Ming Cai Zhouli Li Shuang Wang Xin Hong Bingyi Shi 《Cell biochemistry and biophysics》2013,67(3):1067-1074
Immunotoxins with selective cytotoxicity are frequently used as therapeutic immunosuppressive agents in solid-organ transplantation because of their efficiency and high specificity. In this study, we present a new recombinant immunotoxin termed anti-CTLA-4-scFv–melittin prepared from Escherichia coli aimed at clearing activated T cells at the same time avoiding all-round decline in systematic immunity. This fusion protein is composed of anti-CTLA-4-scFv unit and melittin analog unit with properties of low immunogenicity and selective cytotoxicity to CTLA-4-positive T cells. In preliminary biological activity assays, our results confirmed the feasibility of activated T cell clearance strategy and there were significant differences in cell survival rates between CTLA-4-positive group and control group at all experimental concentrations of the immunotoxin. The selective cytotoxicity, low immunogenicity, and low production cost make it an attractive alternate to traditional immunosuppressants. 相似文献
39.
Xu Liu Ben Chen Lulan Chen Wan-Ting Ren Juan Liu Guoxiang Wang Wei Fan Xin Wang Yun Wang 《PloS one》2013,8(4)
Phenol red is widely used in cell culture as a pH indicator. Recently, it also has been reported to have estrogen-like bioactivity and be capable of promoting cell proliferation in different cell lines. However, the effect of phenol red on primary neuronal culture has never been investigated. By using patch clamp technique, we demonstrated that hippocampal pyramidal neurons cultured in neurobasal medium containing no phenol red had large depolarization-associated epileptiform bursting activities, which were rarely seen in neurons cultured in phenol red-containing medium. Further experiment data indicate that the suppressive effect of the phenol red on the abnormal epileptiform burst neuronal activities was U-shape dose related, with the most effective concentration at 28 µM. In addition, this concentration related inhibitory effect of phenol red on the epileptiform neuronal discharges was mimicked by 17-β-estradiol, an estrogen receptor agonist, and inhibited by ICI-182,780, an estrogen receptor antagonist. Our results suggest that estrogen receptor activation by phenol red in the culture medium prevents formation of abnormal, epileptiform burst activity. These studies highlight the importance of phenol red as estrogen receptor stimulator and cautions of careful use of phenol red in cell culture media. 相似文献
40.
I.CORRELATIONBETWEENL0ESSSECTI0NAND0XYGENIS0TOPESTAGESIntheYuanloessarea,manyscientiststhinkthatthesedimentsarecontinuousandtheclimaticrecordsareperfect.Thisdoesnotappeartobeaccurate.Sedimentationwasdiscontinuousinmanysections.Forexample,manyscientistsarguethatS2corresp0ndstooxygenisot0peStage7(Liu,l985,KuklaandAn,l989;Dingetal.,l990).lnfact,S2includesthreelayersfS2SSl,S2LLlandS2SS2.Dingetal.(l990)reportedthatthereisathickunitofloessbetweenS2SSlandS2SS2intheBaicao… 相似文献